ELISA法检测组织工程产品中残留牛血清蛋白含量

目的探讨定量检测牛血清白蛋白（Bull serum albumin，BSA）酶联免疫试剂盒用于组织工程医疗产品（Medical products of tissue engineering，TEMPs）残留BSA检测的适用性，以及清洗程序对产品BSA残留量的影响。材料和方法检测了三种组织工程医疗产品（组织工程骨、组织工程肌腱以及组织工程角膜贴片）中BSA的残留量，并比较了经不同清洗程序清洗后角膜贴片中残留BSA含量。结果（1）组织工程骨、组织工程肌腱以及组织工程角膜贴片中BSA残留量分别为31．8ng／ml，4．9ng／ml和低于2．5ng／ml，且测定值与其稀释倍数呈正相关；（2）生理盐水及无血清的MEM培养基平均OD值与试剂盒中零浓度标准品接近．表明生理盐水或无血清MEM作为样品浸提介质对最终的实验结果无明显影响；（3）样品复孔比均小于1．24，符合试剂盒有效性要求。经清洗程序2处理后，角膜贴片中残留BSA较清洗程序1明显减少。结论（1）该试剂盒适用于组织工程医疗产品中BSA残留量的检测。（2）对于组织工程医疗产品而言，不同的清洗程序对BSA的残留量有非常明显的影响。生产商应根据其预期用途制定有效的清洗程序，并在说明书中给予足够的阐述。

Quantitative Measure of Residual BSA in TEMPs-ELISA Method

Aim This paper was aimed to study the suitability of a Quantitative measure of residual BSA ELISA kit in detecting residual BSA in TEMPs, and to investigate the effects of rinsing protocol on reducing residual BSA in TEMPs. Materials and methods Three kinds of TEMPs used in this study were： tissue engineered bone, tissue engineered tendon and tissue engineered corneal stroma. The immersion of each product was prepared after the rinsing procedure according to the Direction For Use. Physiological saline and MEM was used as immersion medium. Using a Quantitative measure of residual BSA ELISA kit for BSA, and the results of engineered corneal stroma were compared after different rinsing protocol. Results and conclusion （1）The concentration of BSA residual in the immersions of tissue engineered bone, tissue engineered tendon and tissue engineered corneal stroma was 31.8 ng/ml, 4.9 ng/ml and 〈 2.5 ng/ml, respectively. It also showed some relativity between OD and the sample dilution. The mean value of the OD of physiological saline and MEM were similar to that of 0 ng/ml standard. It indicated that there were no obvious effects on the results when physiological saline or MEM was used as immersion medium. The kit was suitable for detect the residual BSA in TEMPs . （2）The mean value of the OD of the corneal rinsing by protocoll was 2.067, that was upper the reading of the highest standard （40 ng/ml） for 3.3 times, while the OD of that by rinsing protocol 2 was below the reading of the lowest standard （2.5 ng/ml）. The results showed significant different of BSA residual may caused by different rinsing protocol for a TEMPs.